Fig. S1 Pictorial representation of rSt-Phy pPICZαA construct for recombinant P.
pastoris. 5’ AOX1, alcohol oxidase (inducible) promoter sequence; α-factor, secretory
signal sequence; rSt-Phy, recombinant Sporotrichum thermophile phytase; AOX1 TT,
alcohol oxidase transcription termination region; PTEF1, promoter region of
transcription elongation factor 1; PEM7, promoter conferring Zeocin resistance;
Zeocin, a selectable marker; CYC1 TT, cytochrome c1 transcription termination
region; pUC ori, origin of replication
Fig. S2 Q-PCR standard curves for a genomic DNA copies of rSt-Phy and b rSt-PhypPICZαA
100
Relative activity %
80
60
Control
Pepsin
Trypsin
40
20
0
0
10
20
30
40
50
60
Time (min)
Fig. S3 Residual activity of the rSt-Phy, after treating with pepsin (pH 3) and trypsin
(pH 7.5). At same pH rSt-Phy were kept without pepsin and trypsin (control) and there
activity were considered 100 %, relative activity were calculated with control for
pepsin and trypsin respectively at same pH
Table S1 Details of different broiler feed (BPSF, BSF, BFF) ingredients
Feed ingredients
Metabolisable Energy
Crude Protein
Crude Fat
Ash
Fibre
Calcium
Phytate
BPSF
2900 Kcal/kg
~ 22.0 %
~ 2.5 %
10.0 %
~ 4.0 %
0.95%
0.14 - 10.18 %
BSF
BFF
2950 Kcal/kg 3000 Kcal/kg
~ 21.0 %
~18.5 %
~ 2.5 %
~ 3.0 %
6.3 %
5.5 %
3.5 %
3.7 %
1.12 %
0.89 %
0.14 - 10.18 % 0.14 - 10.18 %
Table S2 Comparing codon usage of Pichia pastoris for wild type and codon
optimized gene
Host organism
Codon usage by host for
the wild type gene (St-Phy)
Codon usage by host for
optimized gene (rSt-Phy)
P. pastoris
18 %
52 %